ml of sterilized 10% raffinose and 0.2 ml each of filter-sterilized
M sodium carbonate and 0.32% cobalt chloride (CoCl2·6H2O)
dropwise to each tube. Check pH of one or two tubes; it should be 7.8 ± 0.1. Just before use, steam medium for 10 min; after cooling, add 0.2 ml of filter-sterilized 1.5% sodium ascorbate (prepared daily) to each tube.
M5. Alkaline Peptone Agar Peptone 10 g NaCl 20 g Agar 15 g Distilled water 1 liter
Boil to dissolve ingredients. Adjust pH so that value after sterilization is 8.5 ± 0.2. Autoclave 15 min at 121°C. Solidify agar in tubes as slants.
M6. Alkaline Peptone Salt Broth (APS) Peptone 10 g NaCl 30 g Distilled water 1 liter
Dissolve ingredients. Adjust pH so that value after sterilization is 8.5 ± 0.2. Dispense 10 ml into tubes. Autoclave 10 min at 121°C.
M7. Alkaline Peptone Water Peptone 10 g NaCl 10 g Distilled water l liter